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目的：制备基因工程抗体，减少或消除鼠源性单克隆抗体（ｍＡｂ）在人体内的免疫原性，保留其对人体抗原配体的高度特异性，发展临床导向诊断和治疗。方法：从体外分泌抗转铁蛋白受体ｍＡｂ杂交瘤细胞系７５７９中，对其ｍＡｂ可变区基因进行克隆和序列分析。利用逆转录ＰＣＲ技术扩增轻、重链可变区基因，再分别与ｐＧＥＭＴ载体连接，并克隆于ＪＭ１０９受体菌之中。利用荧光染色链终止法测定其序列，采用ＤＮＡＳＩＳ７分析软件和与ＮＩＨ基因库比较分析。结果：轻、重链可变区分别由１３９和１２８氨基酸残基组成，包括信号肽、互补决定区和骨架氨基酸残基等功能序列。分别属于鼠免疫球蛋白重链Ⅱｃ和κ链Ⅵ家族。结论：来自ｍＡｂ可变区的基因是完整的和具有潜在功能性的，为体外获得抗转铁蛋白受体基因工程抗体奠定了基础。
OBJECTIVE: To prepare a genetically engineered antibody to reduce or eliminate the immunogenicity of murine monoclonal antibody (mAb) in human, retain its high specificity for human antigen ligand and develop clinical oriented diagnosis and treatment. Methods: The mAb variable region gene was cloned and sequenced from in vitro anti-transferrin receptor mAb hybridoma cell line 7579. The light and heavy chain variable region genes were amplified by reverse transcription PCR and ligated with pGEMT vector respectively and cloned into JM109 recipient bacteria. The sequence was determined by fluorescent chain termination method, and the DNASIS7 analysis software and comparative analysis with NIH gene bank were used. RESULTS: The light and heavy chain variable regions consisted of 139 and 128 amino acid residues respectively, including functional sequences of signal peptide, complementarity determining region and backbone amino acid residues. Belong to murine immunoglobulin heavy chain Ⅱ c and κ chain Ⅵ family. CONCLUSION: Genes derived from the mAb variable region are intact and potentially functional and lay the foundation for obtaining genetically engineered antibodies against transferrin receptor in vitro.